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利用RNA测序技术对野生大豆根部NaHCO胁迫响应基因的鉴定与分析

Identification and Analysis of NaHCO Stress Responsive Genes in Wild Soybean () Roots by RNA-seq.

作者信息

Zhang Jinlong, Wang Jiaxue, Jiang Wei, Liu Juge, Yang Songnan, Gai Junyi, Li Yan

机构信息

National Key Laboratory of Crop Genetics and Germplasm Enhancement, National Center for Soybean Improvement, Key Laboratory for Biology and Genetic Improvement of Soybean (Ministry of Agriculture), Jiangsu Collaborative Innovation Center for Modern Crop Production, Nanjing Agricultural University Nanjing, China.

出版信息

Front Plant Sci. 2016 Dec 9;7:1842. doi: 10.3389/fpls.2016.01842. eCollection 2016.

Abstract

Soil alkalinity is a major abiotic constraint to crop productivity and quality. Wild soybean () is considered to be more stress-tolerant than cultivated soybean (), and has considerable genetic variation for increasing alkalinity tolerance of soybean. In this study, we analyzed the transcriptome profile in the roots of an alkalinity tolerant wild soybean variety N24852 at 12 and 24 h after 90 mM NaHCO stress by RNA-sequencing. Compared with the controls, a total of 449 differentially expressed genes (DEGs) were identified, including 95 and 140 up-regulated genes, and 108 and 135 down-regulated genes at 12 and 24 h after NaHCO treatment, respectively. Quantitative RT-PCR analysis of 14 DEGs showed a high consistency with their expression profiles by RNA-sequencing. Gene Ontology (GO) terms related to transcription factors and transporters were significantly enriched in the up-regulated genes at 12 and 24 h after NaHCO stress, respectively. Nuclear factor Y subunit A transcription factors were enriched at 12 h after NaHCO stress, and high percentages of basic helix-loop-helix, ethylene-responsive factor, Trihelix, and zinc finger (C2H2, C3H) transcription factors were found at both 12 and 24 h after NaHCO stress. Genes related to ion transporters such as ABC transporter, aluminum activated malate transporter, glutamate receptor, nitrate transporter/proton dependent oligopeptide family, and S-type anion channel were enriched in up-regulated DEGs at 24 h after NaHCO treatment, implying their roles in maintaining ion homeostasis in soybean roots under alkalinity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed "phenylpropanoid biosynthesis" and "phenylalanine metabolism" pathways might participate in soybean response to alkalinity. This study provides a foundation to further investigate the functions of NaHCO stress-responsive genes and the molecular basis of soybean tolerance to alkalinity.

摘要

土壤碱化是制约作物产量和品质的主要非生物胁迫因素。野生大豆()被认为比栽培大豆()更耐胁迫,并且在提高大豆耐碱性方面具有相当大的遗传变异。在本研究中,我们通过RNA测序分析了耐碱性野生大豆品种N24852在90 mM NaHCO胁迫后12小时和24小时根部的转录组图谱。与对照相比,共鉴定出449个差异表达基因(DEG),其中在NaHCO处理后12小时和24小时分别有95个和140个上调基因,以及108个和135个下调基因。对14个DEG进行的定量RT-PCR分析显示,其表达谱与RNA测序结果高度一致。在NaHCO胁迫后12小时和24小时上调基因中,分别显著富集了与转录因子和转运蛋白相关的基因本体(GO)术语。核因子Y亚基A转录因子在NaHCO胁迫后12小时富集,在NaHCO胁迫后12小时和24小时均发现高比例的碱性螺旋-环-螺旋、乙烯响应因子、三螺旋和锌指(C2H2、C3H)转录因子。与离子转运蛋白相关的基因,如ABC转运蛋白、铝激活苹果酸转运蛋白、谷氨酸受体、硝酸盐转运蛋白/质子依赖性寡肽家族和S型阴离子通道,在NaHCO处理后24小时上调的DEG中富集,这表明它们在碱性条件下维持大豆根部离子稳态中发挥作用。京都基因与基因组百科全书通路富集分析表明,“苯丙烷生物合成”和“苯丙氨酸代谢”通路可能参与大豆对碱化的响应。本研究为进一步研究NaHCO胁迫响应基因的功能以及大豆耐碱性的分子基础提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c603/5161042/dab767ba781e/fpls-07-01842-g001.jpg

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